Figure 1
OC ascites stimulate Mcl-1 protein and mRNA levels. (A) Time course of Mcl-1 protein expression in CaOV3 cells following addition of 10% OVC508 ascites to the cell culture media. Cells were incubated with complete media for 24 h before the addition of ascites. Immunoblots were probed with anti-Mcl-1, anti-Bcl-2, anti-Bcl-XL and anti-tubulin (as a loading control). (B) Immunoblot analysis of Mcl-1, Bcl-2, Bcl-XL and tubulin expression in CaOV3 cells incubated with different OC ascites (10%). Lysates were obtained 2 h following the addition of ascites. (C) Time course of Mcl-1 expression in OVCAR3 cells following addition of 10% OVC508 ascites. (D) Real-time PCR analysis of Mcl-1 transcript levels in the presence or absence of OVC508 ascites (10%) and actinomycin D. Results were standardized using primers of the housekeeping gene RPLPO. Results are expressed as fold change relative to basal levels observed in cells incubated in the absence of ascites. (E) Immunoblot analysis of basal levels of Mcl-1 protein in CaOV3 and OVCAR3 cells. (F) Quantitative real-time PCR of Mcl-1 mRNA expression levels. Expression data were normalized to internal RPLPO RNA expression. Data represents the mean of two experiments ± SEM. ** indicates P < 0.001.